基于高通量RNA-Seq探讨针灸对骨髓抑制小鼠造血微环境的保护及修复作用

Exploring the Protective and Repairing Effects of Acupuncture and Moxibustion on Hem-atopoietic Microenvironment in Myelosuppression Mice Based on High-throughput RNA-Seq

目的探讨针灸参与保护、修复造血微环境的可能作用机制。方法40只雄性KM种小鼠随机分为空白组(Con-trol)、模型组(CTX)、针刺干预组(Acu)、艾灸干预组(Mox),每组10只。CTX组、Acu组、Mox组腹腔注射环磷酰胺制备骨髓抑制模型,于造模后24h开始Acu组予以针刺干预、Mox组予以艾灸干预,Control、CTX组陪同固定,1次/d,连续5d。取各组小鼠骨髓组织提取RNA,进行高通量转录组测序、分析及Real-time PCR实验验证。结果CTX vs Control、Acu vs CTX、Mox vs CTX这3组转录本的交集差异基因有64个,其中具有较高节点热度的中心基因为CXCL12、VEGFA与KDR。与Control组比,CTX组骨髓组织中CXCL12 mRNA表达水平显著降低(P<0.05),CXCR4(CXCL12的受体)、VEGF、VEG-FR-2 mRNA表达水平显著升高(P<0.05);与CTX组比,Acu组、Mox组骨髓组织中CXCL12 mRNA表达水平显著升高(P<0.05),CXCR4、VEGF、VEGFR-2 mRNA表达水平显著下降(P<0.05);与Acu组相比,Mox组CXCL12、CXCR4、VEGF及VEGFR-2 mRNA表达水平无明显差异(P>0.05)。结论针灸缓解骨髓抑制的作用机制,可能与血管新生、正向调节骨髓基质细胞增殖等缓解、修复造血微环境损伤的生物过程有关。

Objective To explore the possible mechanism of acupuncture and moxibustion in protecting and repairing the hemato-poietic microenvironment.Methods 40 male KM mice were randomly divided into Control group,model(CTX)group,acupunc-ture intervention(Acu)group,and moxibustion intervention(Mox)group,with 10 mice in each group.CTX,Acu and Mox groups were injected intraperitoneally with cyclophosphamide to establish bone marrow suppression models.24 hours after model-ing,the Acu group was given acupuncture,the Mox group was given moxibustion,and the Control and CTX groups were accompa-nying fixed,once a day for 5 consecutive days.RNA was extracted from the bone marrow tissue of the mice and were analyzed by High-throughput transcriptome sequencing,analysis and Real-time PCR verification.Results There were 64 common differen-tially expressed genes in the 3 transcripts groups of CTX vs Control,Acu vs CTX,and Mox vs CTX.Among them,the central genes with high node popularity were CXCL12,VEGFA and KDR.Compared with Control group,the expression level of CXCL12 in bone marrow tissue of CTX group was significantly decreased(P<0.05),and the expression levels of CXCR4(the receptor of CXCL12),VEGF and VEGFR-2 were significantly increased(P<0.05).Compared with the Acu group and the Mox group,the CXCL12 expression levels in the bone marrow tissue were significantly increased(P<0.05),and the CXCR4,VEGF,VEG-FR-2 expression levels were significantly decreased(P<0.05);compared with the Acu group,the CXCL12 expression levels in the Mox group,CXCR4,VEGF and VEGFR-2 expression levels had no significant difference(P>0.05).Conclusion The mechanism of acupuncture in alleviating myelosuppression may be related to the biological processes of relieving and repairing hematopoietic microenvironment injury,such as angiogenesis and positive regulation of bone marrow stromal cells proliferation.